title: Zinc-finger-nucleases mediate specific and efficient excision of HIV-1 proviral DNA from infected and latently infected human T cells.
abstract: HIV-infected individuals currently cannot be completely cured because existing antiviral therapy regimens do not address HIV provirus DNA, flanked by long terminal repeats (LTRs), already integrated into host genome. Here, we present a possible alternative therapeutic approach to specifically and directly mediate deletion of the integrated full-length HIV provirus from infected and latently infected human T cell genomes by using specially designed zinc-finger nucleases (ZFNs) to target a sequence within the LTR that is well conserved across all clades. We designed and screened one pair of ZFN to target the highly conserved HIV-1 5'-LTR and 3'-LTR DNA sequences, named ZFN-LTR. We found that ZFN-LTR can specifically target and cleave the full-length HIV-1 proviral DNA in several infected and latently infected cell types and also HIV-1 infected human primary cells in vitro. We observed that the frequency of excision was 45.9% in infected human cell lines after treatment with ZFN-LTR, without significant host-cell genotoxicity. Taken together, our data demonstrate that a single ZFN-LTR pair can specifically and effectively cleave integrated full-length HIV-1 proviral DNA and mediate antiretroviral activity in infected and latently infected cells, suggesting that this strategy could offer a novel approach to eradicate the HIV-1 virus from the infected host in the future.
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China andKey Laboratory of Molecular Virology and Immunology, Institute Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China.
authors.forename: Xiying
authors.initials: X
authors.lastname: Qu
authors.forename: Pengfei
authors.initials: P
authors.lastname: Wang
authors.forename: Donglin
authors.initials: D
authors.lastname: Ding
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.forename: Haibo
authors.initials: H
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Xin
authors.initials: X
authors.lastname: Zhou
authors.forename: Shaohui
authors.initials: S
authors.lastname: Liu
authors.forename: Shiguan
authors.initials: S
authors.lastname: Lin
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Wang
authors.forename: Gongmin
authors.initials: G
authors.lastname: Zhang
authors.forename: Sijie
authors.initials: S
authors.lastname: Liu
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.forename: Jianhua
authors.initials: J
authors.lastname: Wang
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Daru
authors.initials: D
authors.lastname: Lu
authors.forename: Huanzhang
authors.initials: H
authors.lastname: Zhu
mesh_terms: D002478:Cells, Cultured; D004279:DNA, Viral / Q000378:metabolism*; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D011533:Proviruses / Q000235:genetics*; D017384:Sequence Deletion; D013601:T-Lymphocytes / Q000821:virology*; D016662:Virus Integration; D017735:Virus Latency; D016335:Zinc Fingers
chemical_list: D004279:DNA, Viral; D004706:Endodeoxyribonucleases